Human recombinant tissue factor pathway inhibitor induces apoptosis in cultured human endothelial cells.
Tissue factor pathway inhibitor (TFPI) is mainly synthesized in vascular endothelial cells and exhibits a strong and specific inhibitory activity against tissue factor-mediated blood coagulation. In the present study, we demonstrate that human recombinant TFPI (h-rTFPI) inhibits the growth of cultured human umbilical vein endothelial cells (HUVECs) by inducing apoptosis. In a growth-rate assay of HUVECs, the growth of the cultured HUVECs is completely abolished by the addition of 1 microM h-rTFPI to the culture medium containing fetal bovine serum (FBS), basic fibroblast growth factor, and epidermal growth factor. In addition, h-rTFPI and h-rTFPI-C which lacks the carboxyl-terminal basic region prevent the survival of growth-arrested HUVECs which are starved in a medium containing 2%, FBS alone, suggesting that h-rTFPI directly induces the death of these HUVECs. This hypothesis is supported by the finding that h-rTFPI does not inhibit the synthesis of DNA in HUVECs during proliferation, as shown by a 5-bromo-2'-deoxyuridine (BrdU) incorporation assay. Furthermore, Giemsa staining and a gel electrophoretic analysis of DNA fragmentation show that the HUVEC death mediated by h-rTFPI has the typical characteristics of apoptosis. However, the apoptosis in HUVECs is considerably inhibited in the presence of 1 microg/ml of the protein synthesis inhibitor, cycloheximide. Therefore, the process of apoptosis triggered by h-rTFPI is, at least in part, actively conducted by the cells.
Hamuro
T
T
The Chemo-Sero-Therapeutic Research Institute, Kikuchi, Kumamoto, Japan.
Kamikubo
Y
Y
Nakahara
Y
Y
Miyamoto
S
S
Funatsu
A
A
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D017209:Apoptosis; D016466:CHO Cells; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000187:drug effects*; D006131:Growth Inhibitors / Q000494:pharmacology*; D006801:Humans; D008074:Lipoproteins / Q000235:genetics / Q000494:pharmacology*; D011506:Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D006131:Growth Inhibitors; D008074:Lipoproteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; C528024:SHC1 protein, human; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; C051928:lipoprotein-associated coagulation inhibitor